Human/Mouse/Rat DOK7 Antibody

Detection of DOK7 by Western Blot

hDOK7 overexpression stimulates long-term elevation of DOK7 protein expression in mouse hindlimb muscles but not the heart(A) Western blots of tissue lysates probed with anti-DOK7 antibody shown at 55 kDa (middle band). Tissue from three mice each untreated or injected with high- or low-dose AAV9-tMCK-DOK7 was run alongside internal standards (ISs) produced in triplicate, enabling cross-gel comparison. Tissues were taken from mice at 3 months (M) and 6 months of age.(B–D) Bar charts comparing the effect of dose on tissue-specific DOK7 expression. Western blot data were quantified and normalized to the average value of three internal standards. DOK7 expression was significantly increased at 3 and 6 months following high-dose treatment in the hindlimb soleus muscle (B) and lumbricals (C) but not in heart tissue (D), when compared to untreated controls. Specifically, there was a 3.8-fold increase in DOK7 expression in the soleus 3 months following treatment and a 1.9-fold increase at 6 months (B; p < 0.0001 and p = 0.019), while in the lumbricals, there was a 4.7-fold increase in DOK7 expression at 3 months and a 4.6-fold increase at 6 months (C; p = 0.0009 and p = 0.0031) (one-way ANOVAs with Šídák’s multiple comparisons, means of n = 3 per group [with SEM], and each data point represents one animal). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37637210), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of DOK7 by Western Blot

AAV9-tMCK-DOK7 increases DOK7 expression in skeletal muscle(A) Schematic of the AAV9-tMCK-DOK7 plasmid showing the skeletal muscle targeting tMCK promoter & DOK7 transgene.(B–D) DOK7 expression across a range of muscles & organs from 1-month-old mice treated with high dose (5e11 vg), low dose (1.25e11 vg), or untreated with AAV9-tMCK-DOK7. “IS” denotes internal standards run in triplicate used to allow comparison between blots. (B) Western blots where the molecular weight of DOK7 is indicated. Dotted lines illustrate where blot sections have been cropped together. (C) Bar graph of endogenous expression in skeletal muscle & tissue shown by quantified WBs. Values are normalized to the average of three DOK7 internal standards (ISs). Each dot represents a single mouse, & bars represent the mean (with SEM) (n = 3 per bar). (D) DOK7 expression in response to high-dose or low-dose treatments were compared by one-way ANOVA with Holm-Šídák’s multiple comparisons. Each bar represents an average of three animals (with SEM), normalized to internal standard (IS) & then normalized to endogenous protein levels for that tissue. Expression increased by 40x over endogenous levels in the GC (p = 0.0114), & the TA (p = 0.0242), hindlimb lumbricals (p = 0.0074), & TVA (p = 0.013) each showed a marked rise in DOK7. The soleus, which exhibited the highest endogenous levels of DOK7 in skeletal muscle, still expressed ∼4x as much DOK7 following high-dose treatment (p = 0.0032). There was also a small but significant increase in DOK7 expression in the heart following high-dose AAV9-tMCK-DOK7 treatment (p = 0.0018, n = 3 in all groups, one-way ANOVAs with Holm-Šídák’s comparisons). SPC, spinal cord; TVA, transverse abdominis; TA, tibialis anterior; GC, gastrocnemius; ∗p < 0.05, ∗∗p < 0.01. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37637210), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of DOK7 by Western Blot

AAV9-tMCK-DOK7 increases DOK7 expression in skeletal muscle(A) Schematic of the AAV9-tMCK-DOK7 plasmid showing the skeletal muscle targeting tMCK promoter & DOK7 transgene.(B–D) DOK7 expression across a range of muscles & organs from 1-month-old mice treated with high dose (5e11 vg), low dose (1.25e11 vg), or untreated with AAV9-tMCK-DOK7. “IS” denotes internal standards run in triplicate used to allow comparison between blots. (B) Western blots where the molecular weight of DOK7 is indicated. Dotted lines illustrate where blot sections have been cropped together. (C) Bar graph of endogenous expression in skeletal muscle & tissue shown by quantified WBs. Values are normalized to the average of three DOK7 internal standards (ISs). Each dot represents a single mouse, & bars represent the mean (with SEM) (n = 3 per bar). (D) DOK7 expression in response to high-dose or low-dose treatments were compared by one-way ANOVA with Holm-Šídák’s multiple comparisons. Each bar represents an average of three animals (with SEM), normalized to internal standard (IS) & then normalized to endogenous protein levels for that tissue. Expression increased by 40x over endogenous levels in the GC (p = 0.0114), & the TA (p = 0.0242), hindlimb lumbricals (p = 0.0074), & TVA (p = 0.013) each showed a marked rise in DOK7. The soleus, which exhibited the highest endogenous levels of DOK7 in skeletal muscle, still expressed ∼4x as much DOK7 following high-dose treatment (p = 0.0032). There was also a small but significant increase in DOK7 expression in the heart following high-dose AAV9-tMCK-DOK7 treatment (p = 0.0018, n = 3 in all groups, one-way ANOVAs with Holm-Šídák’s comparisons). SPC, spinal cord; TVA, transverse abdominis; TA, tibialis anterior; GC, gastrocnemius; ∗p < 0.05, ∗∗p < 0.01. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37637210), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of DOK7 by Western Blot

hDOK7 overexpression stimulates long-term elevation of DOK7 protein expression in mouse hindlimb muscles but not the heart(A) Western blots of tissue lysates probed with anti-DOK7 antibody shown at 55 kDa (middle band). Tissue from three mice each untreated or injected with high- or low-dose AAV9-tMCK-DOK7 was run alongside internal standards (ISs) produced in triplicate, enabling cross-gel comparison. Tissues were taken from mice at 3 months (M) and 6 months of age.(B–D) Bar charts comparing the effect of dose on tissue-specific DOK7 expression. Western blot data were quantified and normalized to the average value of three internal standards. DOK7 expression was significantly increased at 3 and 6 months following high-dose treatment in the hindlimb soleus muscle (B) and lumbricals (C) but not in heart tissue (D), when compared to untreated controls. Specifically, there was a 3.8-fold increase in DOK7 expression in the soleus 3 months following treatment and a 1.9-fold increase at 6 months (B; p < 0.0001 and p = 0.019), while in the lumbricals, there was a 4.7-fold increase in DOK7 expression at 3 months and a 4.6-fold increase at 6 months (C; p = 0.0009 and p = 0.0031) (one-way ANOVAs with Šídák’s multiple comparisons, means of n = 3 per group [with SEM], and each data point represents one animal). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37637210), licensed under a CC-BY license. Not internally tested by R&D Systems.